大鼠脊髓神经元的分离、培养与鉴定  

Isolation,culture and identification of spinal cord neurons from neonatal rats

作　　者：王斌[1] 郭伟韬[1] 黄云[1] 

机构地区：[1]广东医学院附属医院骨外科,广东湛江524001

出　　处：《海南医学》2012年第6期21-22,共2页Hainan Medical Journal

基　　金：广东省自然科学基金(编号:S2011010004096);广东省医学科研基金(编号:A2010431;A2009477)

摘　　要：目的通过对新生大鼠脊髓神经元分离、培养过程中细节的探讨,建立一种高效、稳定的大鼠脊髓神经元培养方法,为研究脊髓损伤相关的病理生理及其治疗提供种子细胞。方法取新生SD大鼠的脊髓组织,通过木瓜酶消化制成单细胞悬液,经差速贴壁后种于12孔细胞培养板内,用无血清培养基培养,倒置显微镜观察细胞生长状态,免疫细胞化学对神经元β-ⅢTubulin行特异性荧光素染色,结合DAPI核染色鉴定神经细胞及其含量。结果该方法培养的脊髓神经元生长状态良好、密度适中,纯度较高,约83%。结论采用木瓜酶消化、差速贴壁及无血清培养的新生大鼠脊髓神经元符合体外实验的要求,为进一步进行相关实验提供目的细胞。Objective To establish a highly effective and stable method for isolating and cultivating spinal cord neurons, and to provide seed cells for studying the pathophysiolog, abnormal functions and trealmcut of spinal cord injury. Methods Spinal cord tissues of neonatal rats were dissected and cell suspensions were made by papain dissociation. The cell suspensions were cultured in 12 well cell culture cluster with serum-free culture medium after differential adherence, then the morphological development of neurons was observed by invert microscope. The neurons can be identified by immunocytochemistry withfl-m Tubulin. Results The spinal cord neurons had great growth state, appropriate density and high purity, which reached up to about 83%. Conclusion The spinal cord neurons of neonatal rats cultured by papain dissociation, differential adherence and serum-free culture qualify the needs for in vitro experiments.

关 键 词：脊髓 神经元 细胞培养 分离 无血清培养基 

分 类 号：R-332[医药卫生]